Difference between revisions of "Team:Jilin China/InterLab"
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<h2>Introduction</h2> | <h2>Introduction</h2> | ||
| − | <p> | + | <p>Reliable and repeatable measurement remains one of the tough challenges in synthetic biology. In order to overcome this problem, the Measurement Committee, through the InterLab study, has been developing a measurement procedure for green fluorescent protein (GFP) for the last four years. The CFU count is a new feature of interlab study this year. The goal of the 2018 Interlab study is to determine how reliably known parts can conform to expectations and previous results and to calibrate OD¬ to colony forming unit (CFU) counts, which are directly relatable to the cell concentration of the culture, i.e. viable cell counts per mL. The CFU protocol assumes that 1 bacterial cell will give rise to 1 colony.</p> |
| − | + | <p>For a more detailed description visit the homepage of the iGEM's measurement committee.</p> | |
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Revision as of 13:19, 23 September 2018
Interlab
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Bronze Medal Criterion #4
Standard Tracks: Participate in the Interlab Measurement Study and/or obtain new, high quality experimental characterization data for an existing BioBrick Part or Device and enter this information on that part's Main Page in the Registry. The part that you are characterizing must NOT be from a 2018 part number range.
For teams participating in the InterLab study, all work must be shown on this page.
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Introduction
Reliable and repeatable measurement remains one of the tough challenges in synthetic biology. In order to overcome this problem, the Measurement Committee, through the InterLab study, has been developing a measurement procedure for green fluorescent protein (GFP) for the last four years. The CFU count is a new feature of interlab study this year. The goal of the 2018 Interlab study is to determine how reliably known parts can conform to expectations and previous results and to calibrate OD¬ to colony forming unit (CFU) counts, which are directly relatable to the cell concentration of the culture, i.e. viable cell counts per mL. The CFU protocol assumes that 1 bacterial cell will give rise to 1 colony.
For a more detailed description visit the homepage of the iGEM's measurement committee.
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Device We Received
Negative Control BBa_R0040 Positive Control BBa_I20270 Test Device 1 BBa_J364000 Test Device 2 BBa_J364001 Test Device 3 BBa_J364002 Test Device 4 BBa_J364003 Test Device 5 BBa_J364004 Test Device 6 BBa_J364005 -
Protocol
We strictly followed the protocol provided by iGEM. We measured the experiment of OD600 reference point and fitted the fluorescence standard curve first and then detected GFP expression in our E.coli with the plate reader Synergy HT from BioTek.
This is the transformation protocol.
This is the interlab plate reader protocol.
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Raw Data
LUDOX CL-X H2O Replicate 1 0.057 0.038 Replicate 2 0.057 0.039 Replicate 3 0.057 0.039 Replicate 4 0.057 0.041 Arith. Mean 0.057 0.039 Corrected Abs600 0.018 Reference OD600 0.063 OD600/Abs600 3.549 Number of Particles 235294117.647 117647058.824 58823529.412 29411764.706 14705882.353 7352941.176 3676470.588 1838235.294 919117.647 459558.824 229779.412 0.0 Replicate 1 0.703 0.369 0.199 0.141 0.1 0.066 0.051 0.044 0.043 0.041 0.041 0.039 Replicate 2 0.644 0.375 0.219 0.12 0.084 0.064 0.053 0.044 0.041 0.038 0.04 0.039 Replicate 3 0.638 0.359 0.209 0.122 0.08 0.059 0.048 0.044 0.042 0.041 0.043 0.04 Replicate 4 0.651 0.365 0.204 0.123 0.08 0.058 0.047 0.044 0.042 0.042 0.04 0.039 Arith. Mean 0.659 0.367 0.208 0.127 0.086 0.062 0.05 0.044 0.042 0.041 0.041 0.039 Arith. Std.Dev. 0.03 0.007 0.009 0.01 0.01 0.004 0.003 0.0 0.001 0.002 0.001 0.001 Arith. Net Mean 0.62 0.328 0.168 0.087 0.047 0.022 0.011 0.005 0.003 0.001 0.002 Particles / OD Number of Particles 235294117.647 117647058.824 58823529.412 29411764.706 14705882.353 7352941.176 3676470.588 1838235.294 919117.647 459558.824 229779.412 Mean particles / Abs600 379659729.967 358953650.11 349101064.758 337097589.752 314564328.405 326797385.621 350140056.022 386996904.025 334224598.93 367647058.824 131302521.008 Mean of med-high levels: 337302803.729 Fluorescein uM 10.0 5.0 2.5 1.25 0.625 0.312 0.156 0.078 0.039 0.02 0.01 0.0 Replicate 1 77669.0 38692.0 20601.0 10396.0 5058.0 1898.0 1532.0 760.0 382.0 192.0 100.0 10.0 Replicate 2 80872.0 37847.0 20419.0 10215.0 5014.0 2484.0 1212.0 618.0 329.0 155.0 81.0 11.0 Replicate 3 80348.0 39774.0 19819.0 10018.0 4892.0 2221.0 1059.0 522.0 256.0 127.0 65.0 11.0 Replicate 4 78408.0 39810.0 19893.0 10190.0 4987.0 2525.0 1201.0 608.0 309.0 159.0 97.0 12.0 Arith. Mean 79324.25 39030.75 20183.0 10204.75 4987.75 2282.0 1251.0 627.0 319.0 158.25 85.75 11.0 Arith. Std.Dev. 1530.028 944.399 386.012 154.677 70.22 289.269 199.872 98.583 52.083 26.625 16.153 0.816 Arith. Net Mean 79313.25 39019.75 20172.0 10193.75 4976.75 2271.0 1240.0 616.0 308.0 147.25 74.75 Fluorescein/a.u. Fluorescein uM 10.0 5.0 2.5 1.25 0.625 0.312 0.156 0.078 0.039 0.02 0.01 uM Fluorescein/a.u. 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 Mean uM fluorescein / a.u.: 0.0 MEFL / a.u.: 768289204.589 Fluorescence Raw Readings: Hour 0: Neg. Control Pos. Control Device 1 Device 2 Device 3 Device 4 Device 5 Device 6 LB + Chlor (blank) Colony 1, Replicate 1 148.0 293.0 262.0 323.0 149.0 621.0 315.0 199.0 148.0 Colony 1, Replicate 2 149.0 293.0 261.0 318.0 146.0 615.0 307.0 195.0 146.0 Colony 1, Replicate 3 150.0 283.0 258.0 315.0 150.0 611.0 305.0 224.0 145.0 Colony 1, Replicate 4 152.0 290.0 254.0 316.0 149.0 619.0 335.0 223.0 140.0 Colony 2, Replicate 1 153.0 296.0 271.0 298.0 154.0 594.0 310.0 206.0 153.0 Colony 2, Replicate 2 155.0 293.0 259.0 310.0 151.0 580.0 301.0 199.0 147.0 Colony 2, Replicate 3 155.0 290.0 258.0 315.0 148.0 589.0 298.0 208.0 146.0 Colony 2, Replicate 4 154.0 290.0 258.0 310.0 154.0 615.0 313.0 202.0 145.0 Hour 6: Neg. Control Pos. Control Device 1 Device 2 Device 3 Device 4 Device 5 Device 6 LB + Chlor (blank) Colony 1, Replicate 1 174.0 1097.0 766.0 1409.0 192.0 3976.0 794.0 591.0 143.0 Colony 1, Replicate 2 187.0 1161.0 801.0 1486.0 204.0 4135.0 835.0 619.0 144.0 Colony 1, Replicate 3 188.0 1170.0 799.0 1548.0 206.0 4242.0 830.0 624.0 149.0 Colony 1, Replicate 4 195.0 1196.0 850.0 1504.0 201.0 4279.0 836.0 624.0 143.0 Colony 2, Replicate 1 179.0 1115.0 1732.0 2000.0 190.0 4127.0 793.0 603.0 147.0 Colony 2, Replicate 2 187.0 1152.0 1675.0 2071.0 207.0 4478.0 821.0 616.0 149.0 Colony 2, Replicate 3 190.0 1105.0 1734.0 2090.0 203.0 4370.0 823.0 617.0 151.0 Colony 2, Replicate 4 185.0 1157.0 1705.0 2105.0 204.0 4386.0 838.0 624.0 143.0 Abs600 Raw Readings: Hour 0: Neg. Control Pos. Control Device 1 Device 2 Device 3 Device 4 Device 5 Device 6 LB + Chlor (blank) Colony 1, Replicate 1 0.072 0.068 0.059 0.071 0.067 0.06 0.056 0.063 0.045 Colony 1, Replicate 2 0.07 0.069 0.055 0.07 0.067 0.062 0.059 0.067 0.044 Colony 1, Replicate 3 0.071 0.068 0.057 0.07 0.066 0.059 0.056 0.074 0.047 Colony 1, Replicate 4 0.071 0.071 0.057 0.07 0.068 0.06 0.059 0.075 0.048 Colony 2, Replicate 1 0.069 0.068 0.057 0.066 0.067 0.062 0.056 0.065 0.047 Colony 2, Replicate 2 0.07 0.069 0.057 0.067 0.066 0.061 0.056 0.063 0.05 Colony 2, Replicate 3 0.072 0.068 0.055 0.065 0.064 0.061 0.057 0.065 0.043 Colony 2, Replicate 4 0.071 0.071 0.058 0.068 0.064 0.065 0.057 0.063 0.045 Hour 6: Neg. Control Pos. Control Device 1 Device 2 Device 3 Device 4 Device 5 Device 6 LB + Chlor (blank) Colony 1, Replicate 1 0.329 0.288 0.126 0.291 0.301 0.251 0.239 0.267 0.045 Colony 1, Replicate 2 0.332 0.293 0.125 0.298 0.313 0.254 0.25 0.28 0.044 Colony 1, Replicate 3 0.331 0.292 0.125 0.302 0.31 0.256 0.244 0.276 0.047 Colony 1, Replicate 4 0.339 0.296 0.13 0.3 0.309 0.257 0.246 0.278 0.047 Colony 2, Replicate 1 0.308 0.294 0.184 0.313 0.294 0.25 0.248 0.264 0.046 Colony 2, Replicate 2 0.32 0.298 0.186 0.321 0.305 0.266 0.259 0.267 0.045 Colony 2, Replicate 3 0.321 0.289 0.182 0.318 0.299 0.259 0.258 0.268 0.043 Colony 2, Replicate 4 0.314 0.301 0.186 0.323 0.304 0.262 0.26 0.27 0.045